Establishment of Milk ELISA for Anti-Bovine Leukemia Virus Antibody Detection

ABSTRACT Bovine leukemia virus (BLV) is a retrovirus that causes enzootic bovine leukosis in cattle, and Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in cattle. Both diseases are chronic in nature and can lead to the disruption of normal immunological or physiological processes. Cattle are the major reservoir of Shiga toxin–producing Escherichia coli (STEC), a cause of foodborne illness in humans. We tested the hypothesis that cattle infected with BLV or MAP are more likely to shed STEC. We conducted a cross-sectional study during the summers of 2011 and 2012 in 11 Michigan cattle herds. A fecal sample from each animal was collected for STEC culture, and multiplex PCR for stx1, stx2, and eaeA was used to screen suspect colonies for STEC confirmation. Antibody detection enzyme-linked immunosorbent assays for BLV and MAP were used to screen serum from each animal. Flow cytometry was used to quantify the percentage of lymphocytes, monocytes, and neutrophils in a subsample (n =497) of blood samples. Of the animals sampled, 34.9% were BLV positive, 2.7% were MAP positive, and 16% were shedding STEC. Cattle in the dairy herds had a higher frequency of BLV and MAP than did those in beef herds, but more cattle in beef herds were shedding STEC. Neither BLV nor MAP was associated with STEC shedding (P values of 0.6838 and 0.3341, respectively). We also observed no association between STEC status and the percentage of neutrophils (P value of 0.3565), lymphocytes (P value of 0.8422), or the lymphocyte-to-monocyte ratio (P value of 0.1800). Although controlling both BLV and MAP is important for overall herd health and productivity, we found no evidence that controlling BLV and MAP has an impact on STEC shedding in cattle.

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Short communication: Relationship between the level of bovine leukemia virus antibody and provirus in blood and milk of cows from a naturally infected herd

Journal of Dairy Science ◽

10.3168/jds.2015-10813 ◽

2016 ◽

Vol 99 (7) ◽

pp. 5629-5634 ◽

Cited By ~ 7

Author(s):

Juan P. Jaworski ◽

Natalia G. Porta ◽

Geronimo Gutierrez ◽

Romina P. Politzki ◽

Irene Álvarez ◽

...

Keyword(s):

Bovine Leukemia Virus ◽

Short Communication ◽

Leukemia Virus ◽

Infected Herd ◽

Virus Antibody ◽

Communication Relationship

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Using a Herd Profile to Determine Age-Specific Prevalence of Bovine Leukemia Virus in Michigan Dairy Herds

Veterinary Medicine International ◽

10.1155/2012/350374 ◽

2012 ◽

Vol 2012 ◽

pp. 1-5 ◽

Cited By ~ 17

Author(s):

Ronald J. Erskine ◽

Paul C. Bartlett ◽

Todd M. Byrem ◽

Chelsea L. Render ◽

Catherine Febvay ◽

...

Keyword(s):

Bovine Leukemia Virus ◽

Leukemia Virus ◽

Malignant Neoplasm ◽

Dairy Herd ◽

Dairy Herds ◽

Management Plans ◽

Milk Elisa ◽

The Mean ◽

Bovine Leukosis ◽

Simple Average

Enzootic bovine leukosis is a contagious disease of cattle caused by the retrovirus, bovine leukemia virus (BLV) and is the most common cause of malignant neoplasm in cattle. In order to facilitate surveillance of this disease in dairy herds, we developed a method to combine ELISA of milk collected during routine production testing with a prescribed sampling of cows that is independent of the proportion of cows within each lactation. In 113 Michigan dairy herds, milk samples from ten cows in each of the 1st, 2nd, 3rd, and ≥4th lactations were analyzed for anti-Bovine Leukemia Virus (BLV) antibodies by milk ELISA. For each herd, a BLV herd profile (BHP) was calculated as the simple average of the percent of BLV-positive cows within each of the four lactation groups. The mean BHP for all herds was 32.8%, with means of 18.5, 28.8, 39.2, and 44.8% of 1st, 2nd, 3rd, and ≥4th lactation animals infected, respectively. In eight herds, we determined the correlation between the BHP, and true herd prevalence by testing the entire lactating herd (r=0.988, P<0.0001). The BHP allows discrimination of lactation-specific BLV prevalence within a dairy herd, to help identify risk factors and management plans that may be important in transmission of BLV.

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Microplate enzyme-linked immunosorbent assay for bovine leukemia virus antibody.

Journal of Clinical Microbiology ◽

10.1128/jcm.13.1.46-48.1981 ◽

1981 ◽

Vol 13 (1) ◽

pp. 46-48 ◽

Cited By ~ 5

Author(s):

G Poli ◽

A Balsari ◽

A Toniolo ◽

W Ponti ◽

G Vacirca

Keyword(s):

Immunosorbent Assay ◽

Bovine Leukemia Virus ◽

Leukemia Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Virus Antibody

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Comparative diagnostics of Cattle leukemia by ELISA method in test kits of various constructions

Veterinary Medicine: inter-departmental subject scientific collection ◽

10.36016/vm-2019-105-6 ◽

2019 ◽

pp. 32-36

Author(s):

B. T. Stegniy ◽

A. I. Zavgorodniy ◽

S. K. Gorbatenko ◽

O. M. Kornieikov ◽

M. Yu. Stegniy ◽

...

Keyword(s):

False Positive ◽

Bovine Leukemia Virus ◽

Leukemia Virus ◽

Antibody Test ◽

Virus Antibody ◽

Specific Antibodies ◽

Test Kit ◽

Diagnostic Capacity ◽

Cattle Blood ◽

Positive Results

The purpose of the work was to carry out comparative analysis of the positive and negative on leukemia cattle blood sera in ELISA kits of different constructions. Research was carried out using “DIA®-BLV-Ab” kit, in which the reaction had been performed in the indirect ELISA, and “ID Screen® BLV Competition” kit in a competitive format. There were used 15 cattle blood sera for testing, in which antibodies to BLV were confirmed in the ID and the ELISA “Bovine leukemia virus antibody test kit” (IDEXX), as well as 10 positive cattle blood sera confirmed in ID, 10 weak positive sera tested in ID and 10 sera with a weak line of precipitate in ID, 34 negative for leukemia blood sera tested in ID, from which 24 were also tested in the ELISA “Bovine leukemia virus antibody test kit”. The “DIA®-BLV-Ab” kit and “ID Screen® BLV Competition” kit determined positive 25 blood sera with antibodies to BLV, which were positive in ID, and 15 samples were also confirmed in IDEXX test kit. When analyzing 10 sera, that were weak positive in ID, the “DIA®-BLV-Ab” kit determined 8 sera as positive and 2 samples as negative. The “ID Screen® BLV Competition” kit detected specific antibodies to all sera. When analyzing 14 sera with a weak precipitate line in ID, the “DIA®-BLV-Ab” kit determined 9 samples as positive and 5 as negative. The “ID Screen® BLV Competition” determined specific antibodies in 11 samples When analyzing 3 sera, the test result was negative in both ELISA kits. The “DIA®-BLV-Ab” kit determined as negative all 34 sera, which were negative in ID, 24 samples from them were negative in IDEXX test kit. In the “ID Screen® BLV Competition” kit 5 false positive results were received. Studies have shown that both test kits have a high diagnostic capacity and detect antibodies to BLV at different concentrations in all positive sera. The “DIA®-BLV-Ab” kit determined 34 sera as negative, in which specific antibodies were absent, and the “ID Screen® BLV Competition” kit identified 5 samples with a false positive result

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